Soybean variety 95M80

ABSTRACT

According to the invention, there is provided a novel soybean variety, designated 95M80. This invention thus relates to the seeds of soybean variety 95M80, to the plants of soybean 95M80, to plant parts of soybean variety 95M80 and to methods for producing a soybean plant produced by crossing soybean variety 95M80 with another soybean plant, using 95M80 as either the male or the female parent. This invention also relates to methods for introgressing a transgenic or mutant trait into soybean variety 95M80 and to the soybean plants and plant parts produced by those methods. This invention also relates to soybean varieties or breeding varieties and plant parts derived from soybean variety 95M80, to methods for producing other soybean varieties or plant parts derived from soybean variety 95M80 and to the soybean plants, varieties, and their parts derived from use of those methods. This invention further relates to soybean seeds, plants, and plant parts produced by crossing the soybean variety 95M80 with another soybean variety.

FIELD OF INVENTION

This invention is in the field of soybean breeding, specificallyrelating to a soybean variety designated 95M80.

BACKGROUND OF INVENTION

The present invention relates to a new and distinctive soybean variety,designated 95M80 which has been the result of years of careful breedingand selection as part of a soybean breeding program. There are numeroussteps in the development of any novel, desirable plant germplasm. Plantbreeding begins with the analysis and definition of problems andweaknesses of the current germplasm, the establishment of program goals,and the definition of specific breeding objectives. The next step isselection of germplasm that possess the traits to meet the programgoals. The goal is to combine in a single variety an improvedcombination of desirable traits from the parental germplasm. Theseimportant traits may include higher seed yield, resistance to diseasesand insects, tolerance to drought and heat, and better agronomicqualities.

These processes, which lead to the final step of marketing anddistribution, can take from six to twelve years from the time the firstcross is made. Therefore, development of new varieties is atime-consuming process that requires precise forward planning, efficientuse of resources, and a minimum of changes in direction.

Soybean (Glycine max), is an important and valuable field crop. Thus, acontinuing goal of soybean breeders is to develop stable, high yieldingsoybean varieties that are agronomically sound. The reasons for thisgoal are to maximize the amount of grain produced on the land used andto supply food for both animals and humans. To accomplish this goal, thesoybean breeder must select and develop soybean plants that have thetraits that result in superior varieties.

Pioneer soybean research staff creates over 500,000 potential newvarieties each year. Of those new varieties, less than 50 and morecommonly less than 25 are actually selected for commercial use.

The soybean is the world's leading source of vegetable oil and proteinmeal. The oil extracted from soybeans is used for cooking oil,margarine, and salad dressings. Soybean oil is composed of saturated,monounsaturated and polyunsaturated fatty acids. It has a typicalcomposition of 11% palmitic, 4% stearic, 25% oleic, 50% linoleic and 9%linolenic fatty acid content (“Economic Implications of Modified SoybeanTraits Summary Report”, Iowa Soybean Promotion Board & American SoybeanAssociation Special Report 92S, May 1990). Changes in fatty acidcomposition for improved oxidative stability and nutrition areconstantly sought after. Industrial uses of soybean oil which issubjected to further processing include ingredients for paints,plastics, fibers, detergents, cosmetics, and lubricants. Soybean oil maybe split, inter-esterified, sulfurized, epoxidized, polymerized,ethoxylated, or cleaved. Designing and producing soybean oil derivativeswith improved functionality, oliochemistry, is a rapidly growing field.The typical mixture of triglycerides is usually split and separated intopure fatty acids, which are then combined with petroleum-derivedalcohols or acids, nitrogen, sulfonates, chlorine, or with fattyalcohols derived from fats and oils.

Soybean is also used as a food source for both animals and humans.Soybean is widely used as a source of protein for animal feeds forpoultry, swine and cattle. During processing of whole soybeans, thefibrous hull is removed and the oil is extracted. The remaining soybeanmeal is a combination of carbohydrates and approximately 50% protein.

For human consumption soybean meal is made into soybean flour which isprocessed to protein concentrates used for meat extenders or specialtypet foods. Production of edible protein ingredients from soybean offersa healthy, less expensive replacement for animal protein in meats aswell as dairy-type products.

SUMMARY OF INVENTION

According to the invention, there is provided a novel soybean variety,designated 95M80. This invention thus relates to the seeds of soybeanvariety 95M80, to the plants of soybean 95M80, to plant parts of soybeanvariety 95M80 and to methods for producing a soybean plant produced bycrossing soybean variety 95M80 with another soybean plant, using 95M80as either the male or the female parent. This invention also relates tomethods for introgressing a transgenic or mutant trait into soybeanvariety 95M80 and to the soybean plants and plant parts produced bythose methods. This invention also relates to soybean varieties orbreeding varieties and plant parts derived from soybean variety 95M80,to methods for producing other soybean varieties or plant parts derivedfrom soybean variety 95M80 and to the soybean plants, varieties, andtheir parts derived from use of those methods. This invention furtherrelates to soybean seeds, plants, and plant parts produced by crossingthe soybean variety 95M80 with another soybean variety.

Definitions

Certain definitions used in the specification are provided below. Alsoin the examples which follow, a number of terms are used. In order toprovide a clear and consistent understanding of the specification andclaims, including the scope to be given such terms, the followingdefinitions are provided:

ALLELE=any of one or more alternative forms of a genetic sequence. In adiploid cell or organism, the two alleles of a given sequence typicallyoccupy corresponding loci on a pair of homologous chromosomes.

BACKCROSSING=Process in which a breeder crosses a progeny variety backto one of the parental genotypes one or more times.

BREEDING=The genetic manipulation of living organisms.

BREEDING CROSS. A cross to introduce new genetic material into a plantfor the development of a new variety. For example, one could cross plantA with plant B, wherein plant B would be genetically different fromplant A. After the breeding cross, the resulting F1 plants could then beselfed or sibbed for one, two, three or more times (F1, F2, F3, etc.)until a new variety is developed. For clarification, such new varietywould be within a pedigree distance of one breeding cross of plants Aand B. The process described above would be referred to as one breedingcycle.

BU/A=Bushels per Acre. The seed yield in bushels/acre is the actualyield of the grain at harvest.

BSR=Brown Stem Rot Tolerance. This is a visual disease score from 1 to 9comparing all genotypes in a given test. The score is based on leafsymptoms of yellowing and necrosis caused by brown stem rot. A score of9 indicates no symptoms. Visual scores range down to a score of 1 whichindicates severe symptoms of leaf yellowing and necrosis.

CW=Canopy Width. This is visual observation of the canopy width from 1to 9 comparing all genotypes in a given test. The higher the score thebetter the canopy width observed.

CNKR=Stem Canker Tolerance. This is a visual disease score from 1 to 9comparing all genotypes in a given test. The score is based uponpremature plant death. A score of 9 indicates no symptoms, whereas ascore of 1 indicates the entire experimental unit died very early.

COTYLEDON=A cotyledon is a type of seed leaf. The cotyledon contains thefood storage tissues of the seed.

ELITE VARIETY=A variety that is sufficiently homozygous and homogeneousto be used for commercial grain production. An elite variety may also beused in further breeding.

EMBRYO=The embryo is the small plant contained within a mature seed.

EMGSC=Emergence Score. The percentage of emerged plants in a plotrespective to the number of seeds planted.

F3=This symbol denotes a generation resulting from the selfing of the F2generation along with selection for type and rogueing of off-types. The“F” number is a term commonly used in genetics, and designates thenumber of the filial generation. The “F3” generation denotes theoffspring resulting from the selfing or self mating of members of thegeneration having the next lower “F” number, viz. the F2 generation.

FEC=Iron-deficiency Chlorosis. Plants are scored 1 to 9 based on visualobservations. A score of 1 indicates the plants are dead or dying fromiron-deficiency chlorosis, a score of 5 means plants have intermediatehealth with some leaf yellowing and a score of 9 means no stunting ofthe plants or yellowing of the leaves. Plots are usually scored in midJuly.

FECL=Iron-deficiency Chlorosis. Plants are scored 1 to 9 based on visualobservations. A score of 1 indicates the plants are dead or dying fromiron-deficiency chlorosis, a score of 5 means plants have intermediatehealth with some leaf yellowing and a score of 9 means no stunting ofthe plants or yellowing of the leaves. Plots are scored around midAugust.

FEY=Frogeye Tolerance. This is a visual disease score from 1 to 9comparing all genotypes in a given test. The score is based upon leaflesions. A score of 9 indicates no lesions, whereas a score of 1indicates severe leaf necrosis.

GENOTYPE=Refers to the genetic constitution of a cell or organism.

HABIT=This refers to the physical appearance of a plant. It can bedeterminate, semi-determinate, intermediate, or indeterminate. Insoybeans, indeterminate varieties are those in which stem growth is notlimited by formation of a reproductive structure (i.e., flowers, podsand seeds) and hence growth continues throughout flowering and duringpart of pod filling. The main stem will develop and set pods over aprolonged period under favorable conditions. In soybeans, determinatevarieties are those in which stem growth ceases at flowering time. Mostflowers develop simultaneously, and most pods fill at approximately thesame time. The terms semi-determinate and intermediate are also used todescribe plant habit and are defined in Bernard, R. L. 1972. “Two genesaffecting stem termination in soybeans.” Crop Science 12:235–239;Woodworth, C. M. 1932. “Genetics and breeding in the improvement of thesoybean.” Bull. Agric. Exp. Stn. (Illinois) 384:297–404; Woodworth, C.M. 1933. “Genetics of the soybean.” J. Am. Soc. Agron. 25:36–51. skilledin the art, the foregoing are only some of the methods and compositionsthat illustrated the embodiments of the foregoing invention. It will beapparent to those of ordinary skill in the art that variations, changes,modifications and alterations may be applied to the compositions and/ormethods described herein without departing from the true spirit, conceptand scope of the invention.

HERBRES=Herbicide Resistance. This indicates that the plant is moretolerant to the herbicide shown than the level of herbicide toleranceexhibited by wild type plants. A designation of RR indicates toleranceto glyphosate and a designation of STS indicates tolerance tosulfonylurea herbicides.

HGT=Plant Height. Plant height is taken from the top of the soil to toppod of the plant and is measured in inches.

HILUM=This refers to the scar left on the seed which marks the placewhere the seed was attached to the pod prior to it (the seed) beingharvested.

HYPL=Hypocotyl Elongation. This score indicates the ability of the seedto emerge when planted 3″ deep in sand pots and with a controlledtemperature of 25° C. The number of plants that emerge each day arecounted. Based on this data, each genotype is given a 1 to 9 score basedon its rate of emergence and percent of emergence. A score of 9indicates an excellent rate and percent of emergence, an intermediatescore of 5 indicates average ratings and a 1 score indicates a very poorrate and percent of emergence.

HYPOCOTYL=A hypocotyl is the portion of an embryo or seedling betweenthe cotyledons and the root. Therefore, it can be considered atransition zone between shoot and root.

LDGSEV=Lodging Resistance. Lodging is rated on a scale of 1 to 9. Ascore of 9 indicates erect plants. A score of 5 indicates plants areleaning at a 45° angle in relation to the ground and a score of 1indicates plants are laying on the ground.

LEAFLETS=These are part of the plant shoot, and they manufacture foodfor the plant by the process of photosynthesis.

LINKAGE=Refers to a phenomenon wherein alleles on the same chromosometend to segregate together more often than expected by chance if theirtransmission was independent.

LINKAGE DISEQUILIBRIUM=Refers to a phenomenon wherein alleles tend toremain together in linkage groups when segregating from parents tooffspring, with a greater frequency than expected from their individualfrequencies.

LLE=Linoleic Acid Percent. Linoleic acid is one of the five mostabundant fatty acids in soybean seeds. It is measured by gaschromatography and is reported as a percent of the total oil content.

LLN=Linolenic Acid Percent. Linolenic acid is one of the five mostabundant fatty acids in soybean seeds. It is measured by gaschromatography and is reported as a percent of the total oil content.

MAT ABS=Absolute Maturity. This term is defined as the length of timefrom planting to complete physiological development (maturity). Theperiod from planting until maturity is reached is measured in days,usually in comparison to one or more standard varieties. Plants areconsidered mature when 95% of the pods have reached their mature color.

MATURITY GROUP=This refers to an agreed-on industry division of groupsof varieties, based on the zones in which they are adapted primarilyaccording to day length or latitude. They consist of very long daylength varieties (Groups 000, 00, 0), and extend to very short daylength varieties (Groups VII, VIII, IX, X).

OIL=Oil Percent. Soybean seeds contain a considerable amount of oil. Oilis measured by NIR spectrophotometry, and is reported on an as ispercentage basis.

OLC=Oleic Acid Percent. Oleic acid is one of the five most abundantfatty acids in soybean seeds. It is measured by gas chromatography andis reported as a percent of the total oil content.

PEDIGREE DISTANCE=Relationship among generations based on theirancestral links as evidenced in pedigrees. May be measured by thedistance of the pedigree from a given starting point in the ancestry.

PERCENT IDENTITY. Percent identity as used herein refers to thecomparison of the homozygous alleles of two soybean varieties. Percentidentity is determined by comparing a statistically significant numberof the homozygous alleles of two developed varieties. For example, apercent identity of 90% between soybean variety 1 and soybean variety 2means that the two varieties have the same allele at 90% of their loci.

PERCENT SIMILARITY. Percent similarity as used herein refers to thecomparison of the homozygous alleles of a soybean variety such as 95M80with another plant, and if the homozygous allele of 95M80 matches atleast one of the alleles from the other plant then they are scored assimilar. Percent similarity is determined by comparing a statisticallysignificant number of loci and recording the number of loci with similaralleles as a percentage. A percent similarity of 90% between 95M80 andanother plant means that 95M80 matches at least one of the alleles ofthe other plant at 90% of the loci.

PLANT. As used herein, the term “plant” includes reference to animmature or mature whole plant, including a plant from which seed orgrain or anthers have been removed. Seed or embryo that will produce theplant is also considered to be the plant.

PLANT PARTS. As used herein, the term “plant parts” includes leaves,stems, roots, root tips, anthers, seed, grain, embryo, pollen, ovules,flowers, cotyledon, hypocotyl, pod, flower, shoot and stalk, tissue,cells and the like.

PLM=Palmitic Acid Percent. Palmitic acid is one of the five mostabundant fatty acids in soybean seeds. It is measured by gaschromatography and is reported as a percent of the total oil content.

POD=This refers to the fruit of a soybean plant. It consists of the hullor shell (pericarp) and the soybean seeds.

PRT=Phytophthora Tolerance. Tolerance to Phytophthora root rot is ratedon a scale of 1 to 9, with a score of 9 being the best or highesttolerance ranging down to a score of 1 which indicates the plants haveno tolerance to Phytophthora.

PRMMAT=Predicted Relative Maturity. Soybean maturities are divided intorelative maturity groups. In the United States the most common maturitygroups are 00 through VII. Within maturity groups 00 through V aresub-groups. A sub-group is a tenth of a relative maturity group. Withinnarrow comparisons, the difference of a tenth of a relative maturitygroup equates very roughly to a day difference in maturity at harvest.

PRO=Protein Percent. Soybean seeds contain a considerable amount ofprotein. Protein is generally measured by NIR spectrophotometry, and isreported on a dry weight basis.

PUBESCENCE=This refers to a covering of very fine hairs closely arrangedon the leaves, stems and pods of the soybean plant.

RKI=Root-knot Nematode, Southern. This is a visual disease score from 1to 9 comparing all genotypes in a given test. The score is based upondigging plants to visually score the roots for presence or absence ofgalling. A score of 9 indicates that there is no galling of the roots, ascore of 1 indicates large severe galling cover most of the root systemwhich results in pre-mature death from decomposing of the root system.

RKA=Root-knot Nematode, Peanut. This is a visual disease score from 1 to9 comparing all genotypes in a given test. The score is based upondigging plants to look at the roots for presence or absence of galling.A score of 9 indicates that there is no galling of the roots, a score of1 indicates large severe galling cover most of the root system whichresults in pre-mature death from decomposing of the root system.

SCN=Soybean Cyst Nematode Resistance. The score is based on resistanceto a particular race of soybean cyst nematode, such as race 1, 2, 3, 5or 14. Scores are visual observations of resistance as versus othergenotypes in the test, with a higher score indicating a higher level ofresistance.

SD VIG=Seedling Vigor. The score is based on the speed of emergence ofthe plants within a plot relative to other plots within an experiment. Ascore of 9 indicates that 90% of plants growing have expanded firstleaves. A score of 1 indicates no plants have expanded first leaves.

SDS=Sudden Death Syndrome. Tolerance to Sudden Death Syndrome is ratedon a scale of 1 to 9, with a score of 1 being very susceptible rangingup to a score of 9 being tolerant.

S/LB=Seeds per Pound. Soybean seeds vary in seed size, therefore, thenumber of seeds required to make up one pound also varies. This affectsthe pounds of seed required to plant a given area, and can also impactend uses.

SHATTR=Shattering. This refers to the amount of pod dehiscence prior toharvest. Pod dehiscence involves seeds falling from the pods to thesoil. This is a visual score from 1 to 9 comparing all genotypes withina given test. A score of 9 means pods have not opened and no seeds havefallen out. A score of 5 indicates approximately 50% of the pods haveopened, with seeds falling to the ground and a score of 1 indicates 100%of the pods are opened.

SHOOTS=These are a portion of the body of the plant. They consist ofstems, petioles and leaves.

STC=Stearic Acid Percent. Stearic acid is one of the five most abundantfatty acids in soybean seeds. It is measured by gas chromatography andis reported as a percent of the total oil content.

WH MD=White Mold Tolerance. This is a visual disease score from 1 to 9comparing all genotypes in a given test. The score is based uponobservations of mycelial growth and death of plants. A score of 9indicates no symptoms. Visual scores of 1 indicate complete death of theexperimental unit.

Definitions for Area of Adaptability

When referring to area of adaptability, such term is used to describethe location with the environmental conditions that would be well suitedfor this soybean variety. Area of adaptability is based on a number offactors, for example: days to maturity, insect resistance, diseaseresistance, and drought resistance. Area of adaptability does notindicate that the soybean variety will grow in every location within thearea of adaptability or that it will not grow outside the area. Area ofadaptability may also be used to refer to the soil or growingconditions.

-   Midwest: Iowa and Missouri-   Heartland: Illinois and the western half of Indiana-   Plains: ⅔ of the eastern parts of South Dakota and Nebraska-   North Central: Minnesota, Wisconsin, the Upper Peninsula of    Michigan, and the eastern half of North Dakota-   Mideast: Michigan, Ohio, and the eastern half of Indiana-   Eastern: Pennsylvania, Delaware, Maryland, Rhode Island, New Jersey,    Connecticut, Massachusetts, New York, Vermont, and Maine-   Southern: Virginia, West Virginia, Tennessee, Kentucky, Arkansas,    North Carolina, South Carolina, Georgia, Florida, Alabama,    Mississippi, and Louisiana-   Western: Texas, Kansas, Colorado, Oklahoma, New Mexico, Arizona,    Utah, Nevada, California, Washington, Oregon, Montana, Idaho,    Wyoming, the western half of North Dakota, and the western ⅓ South    Dakota and Nebraska-   PMG infested soils: soils containing Phytophthora sojae-   Narrow rows: 7″ and 15″ row spacing-   High yield environments: areas which lack normal stress for example    they have sufficient rainfall, water drainage, low disease pressure,    and low weed pressure-   Tough environments: areas which have stress challenges, opposite of    a high yield environment-   SCN infected soils: soils containing soybean cyst nematode other    areas of adaptation include the soybean growing regions of Canada,    tight clay soils, light sandy soils and no-till locations.

DETAILED DESCRIPTION OF INVENTION

The variety of the invention has shown uniformity and stability for alltraits, as described in the following variety description information.It has been self-pollinated a sufficient number of generations, withcareful attention to uniformity of plant type to ensure a sufficientlevel of homozygosity and phenotypic stability. The variety has beenincreased with continued observation for uniformity. No variant traitshave been observed or are expected.

Soybean variety 95M80 is particularly adapted to the Southern UnitedStates and for use in SCN infected soils.

Soybean variety 95M80 demonstrates a valuable combination of traits,including resistant to labeled rates of glyphosate herbicides,resistance to Race 3 soybean cyst nematode, and excellent resistance tostem canker.

Soybean variety 95M80 exhibits a relative maturity of 5 and a subgroupof approximately 8. A variety description of Soybean variety 95M80 isprovided in Table 1. Traits reported are average values for alllocations and years or samples measured.

Soybean variety 95M80, being substantially homozygous, can be reproducedby planting seeds of the variety, growing the resulting soybean plantsunder self-pollinating or sib-pollinating conditions, and harvesting theresulting seed, using techniques familiar to the agricultural arts.

TABLE 1 Variety Description Information 95M80 PERFORMANCECHARACTERISTICS 95M80 General Characteristics Herbicide Resistance RR,STS RR Avg. Harvest Standability LDGSEV 8 Avg. Field Emergence EMGSC 7Avg. Hypocotyl Length HYPLSC 8 Hypocotyl Length L Avg. Canopy Width (9 =wide) CW 6 Avg. Shattering SHATTR 8 Disease/Insect/Fungal ResistancePhytophthora Race 4 Suscept Phytophthora Race 7 Suscept PhytophthoraRace 25 Suscept Avg. Phytophthora PRT 7 Tolerance Avg. Brown Stem RotBSR Avg. Iron Chlorosis FEC 3 Avg. White Mold WHMD Avg. Cyst NematodeRace 1 SCN1 4 Avg. Cyst Nematode Race 3 SCN3 8 Avg. Cyst Nematode Race 5SCN5 1 Avg. Cyst Nematode Race SCN14 14 Avg. Sudden Death SDS SyndromeAvg. Root-knot Nematode- RKI 6 Southern Avg. Root-knot Nematode- RKA 4Peanut Avg. Stem Canker CNKR 8 Avg. Frogeye Leaf Spot FEY 5 Oil/MealType Avg. Seed Protein (% @ PROT 41.7 Dry Wgt Basis) Avg. Seed Oil (% @Dry OILT 20.4 Wgt Basis) Avg. Seed Size (avg S/LB 3300 seeds/lb) ColorCharacteristics Flower Color FL Purple Pubescence Color PU Gray HilaColor HI Imperfect Black Pod Color PD Tan Seed Coat Luster SCL Dull LeafColor LC Dark Green

PERFORMANCE EXAMPLES OF 95M80

In the examples that follow in Table 2, the traits and characteristicsof soybean variety 95M80 are given in paired comparisons with thePioneer varieties shown in the following tables. Traits reported aremean values for all locations and years where paired comparison data wasobtained.

TABLE 2A VARIETY COMPARISON DATA FOR 95M80 vs. 95B42 YIELD MATABS FECPRTLAB bu/a 60# count HGT in score score OILPCT PROTN G/C countStatistic ABS ABS ABS ABS ABS pct ABS pct ABS ABS 95B42 47.8 133.1 32.96 4.6 18.36 36.21 14.1 95M80 50.4 137.3 32.6 4.7 6.2 17.39 36.34 14.4#Locs 32 15 15 2 4 3 3 2 #Reps 32 15 15 2 4 3 3 2 # Years 2 2 2 1 2 1 11 Abs. Diff 2.6 4.1 0.4 1.3 1.6 0.97 0.13 0.3 Prob 0.002 0.000 0.3570.156 0.003 0.005 0.645 0.814

TABLE 2B VARIETY COMPARISON DATA FOR 95M80 vs. 95B53 YIELD MATABS FECPRTLAB bu/a 60# count HGT in score score OILPCT PROTN G/C countStatistic ABS ABS ABS ABS ABS pct ABS pct ABS ABS 95B53 43.6 135.1 29.95.3 6.8 16.46 37.02 12.9 95M80 50.1 137.6 31.3 4.7 6.3 17.39 36.34 14.3#Locs 40 16 22 2 5 3 3 3 #Reps 40 16 22 2 5 3 3 3 #Years 3 3 3 1 3 1 1 2Abs. Diff 6.6 2.4 1.4 0.7 0.5 0.93 0.68 1.5 Prob 0.000 0.000 0.002 0.5000.135 0.028 0.104 0.101

TABLE 2C VARIETY COMPARISON DATA FOR 95M80 vs. 95B96 YIELD MATABS FECPRTLAB bu/a 60# count HGT in score score OILPCT PROTN G/C countStatistic ABS ABS ABS ABS ABS pct ABS pct ABS ABS 95B96 49.9 139.7 31.86 5.7 17.68 34.71 12.7 95M80 50.1 137.6 31.3 4.7 6.3 17.39 36.34 13.9#Locs 40 16 22 2 5 3 3 4 #Reps 40 16 22 2 5 3 3 4 #Years 3 3 3 1 3 1 1 2Abs. Diff 0.2 2.1 0.5 1.3 0.6 0.29 1.63 1.2 Prob 0.817 0.000 0.173 0.2950.110 0.024 0.028 0.043

TABLE 2D VARIETY COMPARISON DATA FOR 95M80 vs. 96B21 YIELD MATABS FECPRTLAB bu/a 60# count HGT in score score OILPCT PROTN G/C countStatistic ABS ABS ABS ABS ABS pct ABS pct ABS ABS 96B21 51.3 140.4 33.85.8 6 18.46 35.64 15.4 95M80 50.4 137.3 32.6 4.7 6.2 17.39 36.34 14.4#Locs 32 15 15 2 4 3 3 2 #Reps 32 15 15 2 4 3 3 2 #Years 2 2 2 1 2 1 1 1Abs. Diff 0.8 3.1 1.3 1.2 0.1 1.07 0.7 0.9 Prob 0.433 0.000 0.009 0.2580.909 0.022 0.114 0.328

FURTHER EMBODIMENTS OF THE INVENTION

Genetic Marker Profile through SSR and First Generation Progeny

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile which can identify plants of the same variety ora related variety or be used to determine or validate a pedigree.Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asMicrosatellites, and Single Nucleotide Polymorphisms (SNPs). Forexample, see Cregan et. al, “An Integrated Genetic Linkage Map of theSoybean Genome” Crop Science 39:1464–1490 (1999), and Berry et. al.,Assessing Probability of Ancestry Using Simple Sequence Repeat Profiles:Applications to Maize Inbred Lines and Soybean Varieties” Genetics165:331–342 (2003), each of which are incorporated by reference hereinin their entirety.

Particular markers used for these purposes are not limited to anyparticular set of markers, but are envisioned to include any type ofmarker and marker profile which provides a means of distinguishingvarieties. One method of comparison is where only the loci for which95M80 is homozygous are used. For example, one set of publicly availablemarkers which could be used to screen and identify variety 95M80 isdisclosed in Table 3.

TABLE 3 Soybean SSR Marker Set Markers Sctt008 Satt372 Satt495 Satt328Satt582 Satt523 Satt572 Satt389 Satt284 Satt165 Satt543 Satt513 Satt042Satt186 Satt590 Satt300 Sct137 Satt150 Satt050 Satt213 Satt567 Satt385Satt384 Satt540 Satt545 Satt598 Satt175 Satt225 Satt204 Satt551 Satt133Satt602 Satt250 Satt411 Satt452 Satt336 Satt233 Satt193 Satt255 Satt327Satt348 Satt234 Satt421 Satt144 Satt257 Satt470 Sat090 Satt358 Satt455Satt594 Satt259 Satt409 Satt517 Satt420 Satt228 Sat117 Satt262 Sct188Sct187 Satt478 Satt426 Satt353 Satt592 Satt509 Satt568 Satt153 Satt251Sctt009 Satt216 Satt197 Satt279 Satt266 Satt303 Satt367 Satt412 Satt577Satt127 Satt546 Satt467 Sctt012 Satt172 Sct034 Satt270 Sat104 Satt304Satt243 Satt440 Satt601 Satt243 Satt249 Satt556 Satt243 Sct046 Satt122Sct028 Satt596 Satt534 Satt357 Satt380 Satt142 Satt532 Satt183 Satt565Satt221 Satt431 Sct186 Satt383 Satt102 Satt451 Satt295 Satt555 Satt227Satt507 Satt441 Satt432 Satt147 Satt557 Satt457 Satt196 Satt475

Primers and PCR protocols for assaying these and other markers aredisclosed in the Soybase (sponsored by the USDA Agricultural ResearchService and Iowa State University) located at the world wide web at129.186.26.94/SSR.html. In addition to being used for identification ofsoybean variety 95M80 and plant parts and plant cells of variety 95M80,the genetic profile may be used to identify a soybean plant producedthrough the use of 95M80 or to verify a pedigree for progeny plantsproduced through the use of 95M80. The genetic marker profile is alsouseful in breeding and developing backcross conversions.

The present invention comprises a soybean plant characterized bymolecular and physiological data obtained from the representative sampleof said variety deposited with the ATCC. Further provided by theinvention is a soybean plant formed by the combination of the disclosedsoybean plant or plant cell with another soybean plant or cell andcomprising the homozygous alleles of the variety.

Means of performing genetic marker profiles using SSR polymorphisms arewell known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. A markersystem based on SSRs can be highly informative in linkage analysisrelative to other marker systems in that multiple alleles may bepresent. Another advantage of this type of marker is that, through useof flanking primers, detection of SSRs can be achieved, for example, bythe polymerase chain reaction (PCR), thereby eliminating the need forlabor-intensive Southern hybridization. The PCR detection is done by useof two oligonucleotide primers flanking the polymorphic segment ofrepetitive DNA. Repeated cycles of heat denaturation of the DNA followedby annealing of the primers to their complementary sequences at lowtemperatures, and extension of the annealed primers with DNA polymerase,comprise the major part of the methodology.

Following amplification, markers can be scored by gel electrophoresis ofthe amplification products. Scoring of marker genotype is based on thesize of the amplified fragment as measured by base pair weight ormolecular weight of the fragment. While variation in the primer used orin laboratory procedures can affect the reported weight, relative valuesshould remain constant regardless of the specific primer or laboratoryused. When comparing varieties it is preferable if all SSR profiles areperformed in the same lab.

Primers used are publicly available and may be found in the Soybase orCregan supra. See also, PCT Publication No. WO 99/31964 NucleotidePolymorphisms in Soybean, U.S. Pat. No. 6,162,967 Positional Cloning ofSoybean Cyst Nematode Resistance Genes, and US 2002/0129402A1 SoybeanSudden Death Syndrome Resistant Soybeans and Methods of Breeding andIdentifying Resistant Plants, the disclosure of which are incorporatedherein by reference.

The SSR profile of soybean plant 95M80 can be used to identify plantscomprising 95M80 as a parent, since such plants will comprise the samehomozygous alleles as 95M80. Because the soybean variety is essentiallyhomozygous at all relevant loci, most loci should have only one type ofallele present. In contrast, a genetic marker profile of an F1 progenyshould be the sum of those parents, e.g., if one parent was homozygousfor allele x at a particular locus, and the other parent homozygous forallele y at that locus, then the F1 progeny will be xy (heterozygous) atthat locus. Subsequent generations of progeny produced by selection andbreeding are expected to be of genotype x (homozygous), y (homozygous),or xy (heterozygous) for that locus position. When the F1 plant isselfed or sibbed for successive filial generations, the locus should beeither x or y for that position.

In addition, plants and plant parts substantially benefiting from theuse of 95M80 in their development, such as 95M80 comprising a backcrossconversion, transgene, or genetic sterility factor, may be identified byhaving a molecular marker profile with a high percent identity to 95M80.Such a percent identity might be 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9%identical to 95M80.

The SSR profile of 95M80 also can be used to identify essentiallyderived varieties and other progeny varieties developed from the use of95M80, as well as cells and other plant parts thereof. Such plants maybe developed using the markers identified in WO 00/31964, U.S. Pat. No.6,162,967 and US2002/0129402A1. Progeny plants and plant parts producedusing 95M80 may be identified by having a molecular marker profile of atleast 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 79%, 80%,81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99% or 99.5% genetic contribution from soybeanvariety, as measured by either percent identity or percent similarity.Such progeny may be further characterized as being within a pedigreedistance of 95M80, such as within 1,2,3,4 or 5 or lesscross-pollinations to a soybean plant other than 95M80 or a plant thathas 95M80 as a progenitor. Unique molecular profiles may be identifiedwith other molecular tools such as SNPs and RFLPs.

While determining the SSR genetic marker profile of the plants describedsupra, several unique SSR profiles may also be identified which did notappear in either parent of such plant. Such unique SSR profiles mayarise during the breeding process from recombination or mutation. Acombination of several unique alleles provides a means of identifying aplant variety, an F1 progeny produced from such variety, and progenyproduced from such variety.

Introduction of a New Trait or Locus into 95M80

Variety 95M80 represents a new base genetic variety into which a newlocus or trait may be introgressed. Direct transformation andbackcrossing represent two important methods that can be used toaccomplish such an introgression. The term backcross conversion andsingle locus conversion are used interchangeably to designate theproduct of a backcrossing program.

Backcross Conversions of 95M80

A backcross conversion of 95M80 occurs when DNA sequences are introducedthrough backcrossing (Hallauer et al, 1988), with 95M80 utilized as therecurrent parent. Both naturally occurring and transgenic DNA sequencesmay be introduced through backcrossing techniques. A backcrossconversion may produce a plant with a trait or locus conversion in atleast two or more backcrosses, including at least 2 crosses, at least 3crosses, at least 4 crosses, at least 5 crosses and the like. Molecularmarker assisted breeding or selection may be utilized to reduce thenumber of backcrosses necessary to achieve the backcross conversion. Forexample, see Openshaw, S. J. et al., Marker-assisted Selection inBackcross Breeding. In: Proceedings Symposium of the Analysis ofMolecular Data, August 1994, Crop Science Society of America, Corvallis,Oreg., where it is demonstrated that a backcross conversion can be madein as few as two backcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes as vs.unlinked genes), the level of expression of the trait, the type ofinheritance (cytoplasmic or nuclear) and the types of parents includedin the cross. It is understood by those of ordinary skill in the artthat for single gene traits that are relatively easy to classify, thebackcross method is effective and relatively easy to manage. (SeeHallauer et al. in Corn and Corn Improvement, Sprague and Dudley, ThirdEd. 1998). Desired traits that may be transferred through backcrossconversion include, but are not limited to, sterility (nuclear andcytoplasmic), fertility restoration, nutritional enhancements, droughttolerance, nitrogen utilization, altered fatty acid profile, lowphytate, industrial enhancements, disease resistance (bacterial, fungalor viral), insect resistance and herbicide resistance. In addition, anintrogression site itself, such as an FRT site, Lox site or other sitespecific integration site, may be inserted by backcrossing and utilizedfor direct insertion of one or more genes of interest into a specificplant variety. In some embodiments of the invention, the number of locithat may be backcrossed into 95M80 is at least 1, 2, 3, 4, or 5 and/orno more than 6, 5, 4, 3, or 2. A single locus may contain severaltransgenes, such as a transgene for disease resistance that, in the sameexpression vector, also contains a transgene for herbicide resistance.The gene for herbicide resistance may be used as a selectable markerand/or as a phenotypic trait. A single locus conversion of site specificintegration system allows for the integration of multiple genes at theconverted loci.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selfing thefirst backcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the locusof interest. The last backcross generation is usually selfed to givepure breeding progeny for the gene(s) being transferred, although abackcross conversion with a stably introgressed trait may also bemaintained by further backcrossing to the recurrent parent withselection for the converted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype of the recurrent parent. The backcross is a form ofinbreeding, and the features of the recurrent parent are automaticallyrecovered after successive backcrosses. Poehlman, Breeding Field Crops,P. 204 (1987). Poehlman suggests from one to four or more backcrosses,but as noted above, the number of backcrosses necessary can be reducedwith the use of molecular markers. Other factors, such as a geneticallysimilar donor parent, may also reduce the number of backcrossesnecessary. As noted by Poehlman, backcrossing is easiest for simplyinherited, dominant and easily recognized traits.

One process for adding or modifying a trait or locus in soybean variety95M80 comprises crossing 95M80 plants grown from 95M80 seed with plantsof another soybean variety that comprise the desired trait or locus,selecting F1 progeny plants that comprise the desired trait or locus toproduce selected F1 progeny plants, crossing the selected progeny plantswith the 95M80 plants to produce backcross progeny plants, selecting forbackcross progeny plants that have the desired trait or locus and themorphological characteristics of soybean variety 95M80 to produceselected backcross progeny plants; and backcrossing to 95M80 three ormore times in succession to produce selected fourth or higher backcrossprogeny plants that comprise said trait or locus. The modified 95M80 maybe further characterized as having the physiological and morphologicalcharacteristics of soybean variety 95M80 listed in Table 1 as determinedat the 5% significance level when grown in the same environmentalconditions and/or may be characterized by percent similarity or identityto 95M80 as determined by SSR markers. The above method may be utilizedwith fewer backcrosses in appropriate situations, such as when the donorparent is highly related or markers are used in the selection step.Desired traits that may be used include those nucleic acids known in theart, some of which are listed herein, that will affect traits throughnucleic acid expression or inhibition. Desired loci include theintrogression of FRT, Lox and other sites for site specific integration,which may also affect a desired trait if a functional nucleic acid isinserted at the integration site.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny soybean seed byadding a step at the end of the process that comprises crossing 95M80with the introgressed trait or locus with a different soybean plant andharvesting the resultant first generation progeny soybean seed.

Transformation

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions, such as encoding specific protein products. Scientists in thefield of plant biology developed a strong interest in engineering thegenome of plants to contain and express foreign genetic elements, oradditional, or modified versions of native or endogenous geneticelements in order to alter the traits of a plant in a specific manner.Any DNA sequences, whether from a different species or from the samespecies, that are inserted into the genome using transformation arereferred to herein collectively as “transgenes”. In some embodiments ofthe invention, a transformed variant of 95M80 may contain at least onetransgene but could contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10and/or no more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2.Over the last fifteen to twenty years several methods for producingtransgenic plants have been developed, and the present invention alsorelates to transformed versions of the claimed soybean variety 95M80.

One embodiment of the invention is a process for producing soybeanvariety 95M80 further comprising a desired trait, said processcomprising transforming a soybean plant of variety 95M80 with atransgene that confers a desired trait. Another embodiment is theproduct produced by this process. In one embodiment the desired traitmay be one or more of herbicide resistance, insect resistance, diseaseresistance, decreased phytate, or modified fatty acid or carbohydratemetabolism. The specific gene may be any known in the art or listedherein, including; a polynucleotide conferring resistance toimidazolinone, sulfonylurea, glyphosate, glufosinate, triazine andbenzonitrile; a polynucleotide encoding a bacillus thuringiensispolypeptide, a polynucleotide encoding phytase, FAD-2, FAD-3, galactinolsynthase or a raffinose synthetic enzyme; or a polynucleotide conferringresistance to soybean cyst nematode, brown stem rot, phytophthora rootrot, soybean mosaic virus or sudden death syndrome.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67–88 and Armstrong, “The First Decade of Maize Transformation: A Reviewand Future Perspective” (Maydica 44:101–109, 1999). In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Gruber et al., “Vectors for Plant Transformation” in Methods inPlant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J.E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89–119.

The most prevalent types of plant transformation involve theconstruction of an expression vector. Such a vector comprises a DNAsequence that contains a gene under the control of or operatively linkedto a regulatory element, for example a promoter. The vector may containone or more genes and one or more regulatory elements.

A genetic trait which has been engineered into the genome of aparticular soybean plant using transformation techniques, could be movedinto the genome of another variety using traditional breeding techniquesthat are well known in the plant breeding arts. For example, abackcrossing approach may be used to move a transgene from a transformedsoybean variety into an already developed soybean variety, and theresulting backcross conversion plant would then comprise thetransgene(s).

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include, but are not limited to genes;coding sequences; inducible, constitutive, and tissue specificpromoters; enhancing sequences; and signal and targeting sequences. Forexample, see the traits, genes and transformation methods listed in U.S.Pat. No. 6,118,055.

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants that areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, Anal. Biochem. 114: 92–6(1981).

A genetic map can be generated, primarily via conventional RestrictionFragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR)analysis, Simple Sequence Repeats (SSR) and Single NucleotidePolymorphisms (SNP) that identifies the approximate chromosomal locationof the integrated DNA molecule. For exemplary methodologies in thisregard, see Glick and Thompson, METHODS IN PLANT MOLECULAR BIOLOGY ANDBIOTECHNOLOGY 269–284 (CRC Press, Boca Raton, 1993).

Wang et al. discuss “Large Scale Identification, Mapping and Genotypingof Single-Nucleotide Polymorphisms in the Human Genome”, Science,280:1077–1082, 1998, and similar capabilities are becoming increasinglyavailable for the soybean genome. Map information concerning chromosomallocation is useful for proprietary protection of a subject transgenicplant. If unauthorized propagation is undertaken and crosses made withother germplasm, the map of the integration region can be compared tosimilar maps for suspect plants to determine if the latter have a commonparentage with the subject plant. Map comparisons would involvehybridizations, RFLP, PCR, SSR and sequencing, all of which areconventional techniques. SNPs may also be used alone or in combinationwith other techniques.

Likewise, by means of the present invention, plants can be geneticallyengineered to express various phenotypes of agronomic interest. Throughthe transformation of soybean the expression of genes can be modulatedto enhance disease resistance, insect resistance, herbicide resistance,agronomic, grain quality and other traits. Transformation can also beused to insert DNA sequences which control or help controlmale-sterility. DNA sequences native to soybean as well as non-nativeDNA sequences can be transformed into soybean and used to modulatelevels of native or non-native proteins. Various promoters, targetingsequences, enhancing sequences, and other DNA sequences can be insertedinto the genome for the purpose of modulating the expression ofproteins. Reduction of the activity of specific genes (also known asgene silencing, or gene suppression) is desirable for several aspects ofgenetic engineering in plants.

Many techniques for gene silencing are well known to one of skill in theart, including but not limited to antisense technology (see, e.g.,Sheehy et al. (1988) PNAS USA 85:8805–8809; and U.S. Pat. Nos.5,107,065; 5,453,566; and 5,759,829); co-suppression (e.g., Taylor(1997) Plant Cell 9:1245; Jorgensen (1990) Trends Biotech. 8(12):340–344; Flavell (1994) PNAS USA 91:3490–3496; Finnegan et al. (1994)Bio/Technology 12: 883–888; and Neuhuber et al. (1994) Mol. Gen. Genet.244:230–241); RNA interference (Napoli et al. (1990) Plant Cell2:279–289; U.S. Pat. No. 5,034,323; Sharp (1999) Genes Dev. 13:139–141;Zamore et al. (2000) Cell 101:25–33; and Montgomery et al. (1998) PNASUSA 95:15502–15507), virus-induced gene silencing (Burton, et al. (2000)Plant Cell 12:691–705; and Baulcombe (1999) Curr. Op. Plant Bio.2:109–113); target-RNA-specific ribozymes (Haseloff et al. (1988) Nature334: 585–591); hairpin structures (Smith et al. (2000) Nature407:319–320; WO 99/53050; and WO 98/53083); ribozymes (Steinecke et al.((1992) EMBO J. 11:1525; and Perriman et al. ((1993) Antisense Res. Dev.3:253); oligonucleotide mediated targeted modification (e.g., WO03/076574 and WO 99/25853); Zn-finger targeted molecules (e.g., WO01/52620; WO 03/048345; and WO 00/42219); and other methods orcombinations of the above methods known to those of skill in the art.

Exemplary transgenes useful for genetic engineering include, but are notlimited to, those categorized below.

1. Transgenes that Confer Resistance to Insects or Disease and thatEncode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones et al., Science 266: 789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262: 1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinoset al., Cell 78: 1089 (1994) (Arabidopsis RSP2 gene for resistance toPseudomonas syringae). A plant resistant to a disease is one that ismore resistant to a pathogen as compared to the wild type plant.

(B) A Bacillus thuringiensis protein, a derivative thereof or asynthetic polypeptide modeled thereon. See, for example, Geiser et al.,Gene 48: 109 (1986), who disclose the cloning and nucleotide sequence ofa Bt delta-endotoxin gene. Moreover, DNA molecules encodingdelta-endotoxin genes can be purchased from American Type CultureCollection (Rockville, Md.), for example, under ATCC Accession Nos.40098, 67136, 31995 and 31998. Other examples of Bacillus thuringiensistransgenes being genetically engineered are given in the followingpatents and patent applications and hereby are incorporated by referencefor this purpose: U.S. Pat. Nos. 5,188,960; 5,689,052; 5,880,275; WO91/114778; WO 99/31248; WO 01/12731; WO 99/24581; WO 97/40162 and U.S.application Ser. Nos. 10/032,717; 10/414,637; and 10/606,320.

(C) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344: 458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

(D) An insect-specific peptide which, upon expression, disrupts thephysiology of the affected pest. For example, see the disclosures ofRegan, J. Biol. Chem. 269: 9 (1994) (expression cloning yields DNAcoding for insect diuretic hormone receptor), and Pratt et al., Biochem.Biophys. Res. Comm. 163: 1243 (1989) (an allostatin is identified inDiploptera puntata). See also U.S. Pat. No. 5,266,317 to Tomalski etal., who disclose genes encoding insect-specific toxins.

(E) An enzyme responsible for an hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(F) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23: 691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al.,Plant Molec. Biol. 21: 673 (1993), who provide the nucleotide sequenceof the parsley ubi4-2 polyubiquitin gene.

(G) A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24: 757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess etal., Plant Physiol. 104: 1467 (1994), who provide the nucleotidesequence of a maize calmodulin cDNA clone.

(H) A hydrophobic moment peptide. See PCT application WO 95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference for this purpose.

(I) A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure by Jaynes et al., Plant Sci. 89: 43 (1993),of heterologous expression of a cecropin-beta lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

(J) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. Rev. Phytopathol.28: 451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

(K) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et a/., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULARPLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

(L) A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366: 469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

(M) A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo alpha-1,4-D-polygalacturonasesfacilitate fungal colonization and plant nutrient release bysolubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See Lamb etal., Bio/Technology 10: 1436 (1992). The cloning and characterization ofa gene which encodes a bean endopolygalacturonase-inhibiting protein isdescribed by Toubart et al., Plant J. 2: 367 (1992).

(N) A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10: 305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

(O) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, S., Current Biology, 5(2)(1995).

(P) Antifungal genes (Cornelissen and Melchers, Pl. Physiol.101:709–712, (1993) and Parijs et al., Planta 183:258–264, (1991) andBushnell et al., Can. J. of Plant Path. 20(2):137–149 (1998).

(Q) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see U.S. Pat. No. 5,792,931.

(R) Cystatin and cysteine proteinase inhibitors.

(S) Defensin genes. See WO 03000863.

(T) Genes conferring resistance to nematodes, and in particular soybeancyst nematodes. See e.g. PCT Application WO96/30517; PCT ApplicationWO93/19181, WO 03/033651 and Urwin et. al., Planta 204:472–479 (1998).

(U) Genes that confer resistance to Phytophthora Root Rot, such as theRps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2, Rps3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes.See, for example, Shoemaker et al, Phytophthora Root Rot Resistance GeneMapping in Soybean, Plant Genome IV Conference, San Diego, Calif.(1995).

(V) Genes that confer resistance to Brown Stem Rot, such as described inU.S. Pat. No. 5,689,035 and incorporated by reference for this purpose.

2. Transgenes that Confer Resistance to a Herbicide, for Example:

(A) A herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7: 1241 (1988), and Miki et al., Theor. Appl. Genet. 80: 449(1990), respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659;5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;5,928,937; and 5,378,824; and international publication WO 96/33270,which are incorporated herein by reference for this purpose.

(B) Glyphosate (resistance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase (PAT) and Streptomyceshygroscopicus phosphinothricin acetyl transferase (bar) genes), andpyridinoxy or phenoxy proprionic acids and cycloshexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah et al., which discloses the nucleotide sequence of a form of EPSPSwhich can confer glyphosate resistance. U.S. Pat. No. 5,627,061 to Barryet al. also describes genes encoding EPSPS enzymes. See also U.S. Pat.Nos. 6,248,876 B1; 6,040,497; 5,804,425; 5,633,435; 5,145,783;4,971,908; 5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,114 B1;6,130,366; 5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re.36,449; RE 37,287 E; and 5,491,288; and international publications WO97/04103; WO 97/04114; WO 00/66746; WO 01/66704; WO 00/66747 and WO00/66748, which are incorporated herein by reference for this purpose.Glyphosate resistance is also imparted to plants that express a genethat encodes a glyphosate oxido-reductase enzyme as described more fullyin U.S. Pat. Nos. 5,776,760 and 5,463,175, which are incorporated hereinby reference for this purpose. In addition glyphosate resistance can beimparted to plants by the over expression of genes encoding glyphosateN-acetyltransferase. See, for example, U.S. Application Ser. Nos.60/244,385; 60/377,175 and 60/377,719. A DNA molecule encoding a mutantaroA gene can be obtained under ATCC accession No. 39256, and thenucleotide sequence of the mutant gene is disclosed in U.S. Pat. No.4,769,061 to Comai. European patent application No. 0 333 033 to Kumadaet al. and U.S. Pat. No. 4,975,374 to Goodman et al. disclose nucleotidesequences of glutamine synthetase genes which confer resistance toherbicides such as L-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in European PatentNo. 0 242 246 and 0 242 236 to Leemans et al. De Greef et al.,Bio/Technology 7: 61 (1989), describe the production of transgenicplants that express chimeric bar genes coding for phosphinothricinacetyl transferase activity. See also, U.S. Pat. Nos. 5,969,213;5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477;5,646,024; 6,177,616 B1; and 5,879,903, which are incorporated herein byreference for this purpose. Exemplary genes conferring resistance tophenoxy proprionic acids and cycloshexones, such as sethoxydim andhaloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3 genes described byMarshall et al., Theor. Appl. Genet. 83: 435 (1992).

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+genes) and a benzonitrile (nitrilase gene). Przibilla et al.,Plant Cell 3: 169 (1991), describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNos. 53435, 67441 and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J. 285:173 (1992).

(D) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants (see, e.g., Hattori et al. (1995)Mol Gen Genet 246:419). Other genes that confer tolerance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota et al. (1994)Plant PhysiolPlant Physiol 106:17), genes for glutathione reductase andsuperoxide dismutase (Aono et al. (1995) Plant Cell Physiol 36:1687, andgenes for various phosphotransferases (Datta et al. (1992) Plant MolBiol 20:619).

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1;and 5,767,373; and international publication WO 01/12825, which areincorporated herein by reference for this purpose.

3. Transgenes that Confer or Contribute to a Grain Trait, such as:

(A) Modified fatty acid metabolism, for example, by

-   -   (1) Transforming a plant with an antisense gene of stearoyl-ACP        desaturase to increase stearic acid content of the plant. See        Knultzon et al., Proc. Natl. Acad. Sci. USA 89: 2624 (1992),    -   (2) Elevating oleic acid via FAD-2 gene modification and/or        decreasing linolenic acid via FAD-3 gene modification (see U.S.        Pat. Nos. 6,063,947; 6,323,392; and WO 93/11245),    -   (3) Altering conjugated linolenic or linoleic acid content, such        as in WO 01/12800,    -   (4) Modifying LEC1, AGP, Dek1, Superal1, and/or thioredoxin. For        example, see WO 02/42424, WO 98/22604, WO 03/011015, U.S. Pat.        No. 6,423,886 and Rivera-Madrid, R. et. al. Proc. Natl. Acad.        Sci. 92:5620–5624 (1995).

(B) Decreased phytate content, for example, by the

-   -   (1) Introduction of a phytase-encoding gene would enhance        breakdown of phytate, adding more free phosphate to the        transformed plant. For example, see Van Hartingsveldt et al.,        Gene 127: 87 (1993), for a disclosure of the nucleotide sequence        of an Aspergillus niger phytase gene.    -   (2) Introduction of a gene that reduces phytate content. In        maize, this, for example, could be accomplished, by cloning and        then re-introducing DNA associated with one or more of the        alleles, such as the LPA alleles, identified in maize mutants        characterized by low levels of phytic acid, such as in Raboy et        al., Maydica 35: 383 (1990) and/or by altering inositol kinase        activity as in WO 02/059324, US2003/0009011, WO 03/027243,        US2003/0079247 and WO 99/05298.

(C) Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteriol. 170: 810(1988) (nucleotide sequence of Streptococcus mutans fructosyltransferasegene), Steinmetz et al., Mol. Gen. Genet. 200: 220 (1985) (nucleotidesequence of Bacillus subtilis levansucrase gene), Pen et al.,Bio/Technology 10: 292 (1992) (production of transgenic plants thatexpress Bacillus licheniformis alpha-amylase), Elliot et al., PlantMolec. Biol. 21: 515 (1993) (nucleotide sequences of tomato invertasegenes), Søgaard et al., J. Biol. Chem. 268: 22480 (1993) (site-directedmutagenesis of barley alpha-amylase gene), and Fisher et al., PlantPhysiol. 102: 1045 (1993) (maize endosperm starch branching enzyme II).The fatty acid modification genes mentioned above may also be used toeffect starch content and/or composition through the interrelationshipof the starch and oil pathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see WO 00/68393 involving themanipulation of antioxidant levels through alteration of a phytl prenyltransferase and WO 03/082899 through alteration of a homogentisategeranyl geranyl transferase.

(E) Improved digestibility and/or starch extraction through modificationof UDP-D-xylose 4-epimerase, Fragile 1 and 2, Ref1, HCHL, C4H, such asin WO 99/10498.

4. Genes that Control Male-Sterility

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac-PPT (WO 01/29237).

(B) Introduction of various stamen-specific promoters (WO 92/13956, WO92/13957).

(C) Introduction of the barnase and the barstar gene (Paul et al. PlantMol. Biol. 19:611–622, 1992).

5. Genes that create a site for site specific DNA integration. Thisincludes the introduction of FRT sites that may be used in the FLP/FRTsystem and/or Lox sites that may be used in the Cre/Loxp system. Forexample, see Lyznik, et al., Site-Specific Recombination for GeneticEngineering in Plants, Plant Cell Rep (2003) 21:925–932 and WO 99/25821,which are hereby incorporated by reference. Other systems that may beused include the Gin recombinase of phage Mu (Maeser et al., 1991), thePin recombinase of E. coli (Enomoto et al., 1983), and the R/RS systemof the pSR1 plasmid (Araki et al., 1992).6. Genes that affect growth characteristics, such as drought toleranceand nitrogen utilization. For example, see WO 00/73475 where water useefficiency is modulated through alteration of malate.Using 95M80 to Develop other Soybean Varieties

Soybean varieties such as 95M80 are typically developed for use in seedand grain production. However, soybean varieties such as 95M80 alsoprovide a source of breeding material that may be used to develop newsoybean varieties. Plant breeding techniques known in the art and usedin a soybean plant breeding program include, but are not limited to,recurrent selection, mass selection, bulk selection, mass selection,backcrossing, pedigree breeding, open pollination breeding, restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, and transformation. Oftencombinations of these techniques are used. The development of soybeanvarieties in a plant breeding program requires, in general, thedevelopment and evaluation of homozygous varieties. There are manyanalytical methods available to evaluate a new variety. The oldest andmost traditional method of analysis is the observation of phenotypictraits but genotypic analysis may also be used.

Using 95M80 in a Breeding Program

This invention is directed to methods for producing a soybean plant bycrossing a first parent soybean plant with a second parent soybean plantwherein either the first or second parent soybean plant is variety95M80. The other parent may be any other soybean plant, such as asoybean plant that is part of a synthetic or natural population. Anysuch methods using soybean variety 95M80 are part of this invention:selfing, sibbing, backcrosses, mass selection, pedigree breeding, bulkselection, hybrid production, crosses to populations, and the like.These methods are well known in the art and some of the more commonlyused breeding methods are described below. Descriptions of breedingmethods can be found in one of several reference books (e.g., Allard,Principles of Plant Breeding, 1960; Simmonds, Principles of CropImprovement, 1979; Sneep et al., 1979; Fehr, “Breeding Methods forCultivar Development”, Chapter 7, Soybean Improvement, Production andUses, 2^(nd) ed., Wilcox editor, 1987).

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such as95M80 and another soybean variety having one or more desirablecharacteristics that is lacking or which complements 95M80. If the twooriginal parents do not provide all the desired characteristics, othersources can be included in the breeding population. In the pedigreemethod, superior plants are selfed and selected in successive filialgenerations. In the succeeding filial generations the heterozygouscondition gives way to homogeneous varieties as a result ofself-pollination and selection. Typically in the pedigree method ofbreeding, five or more successive filial generations of selfing andselection is practiced: F1→F2; F2→F3; F3→F4; F4→F₅, etc. After asufficient amount of inbreeding, successive filial generations willserve to increase seed of the developed variety. Preferably, thedeveloped variety comprises homozygous alleles at about 95% or more ofits loci.

In addition to being used to create a backcross conversion, backcrossingcan also be used in combination with pedigree breeding. As discussedpreviously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety, the donor parent, to adeveloped variety called the recurrent parent, which has overall goodagronomic characteristics yet lacks that desirable trait or traits.However, the same procedure can be used to move the progeny toward thegenotype of the recurrent parent but at the same time retain manycomponents of the non-recurrent parent by stopping the backcrossing atan early stage and proceeding with selfing and selection. For example, asoybean variety may be crossed with another variety to produce a firstgeneration progeny plant. The first generation progeny plant may then bebackcrossed to one of its parent varieties to create a BC1 or BC2.Progeny are selfed and selected so that the newly developed variety hasmany of the attributes of the recurrent parent and yet several of thedesired attributes of the non-recurrent parent. This approach leveragesthe value and strengths of the recurrent parent for use in new soybeanvarieties.

Therefore, an embodiment of this invention is a method of making abackcross conversion of soybean variety 95M80, comprising the steps ofcrossing a plant of soybean variety 95M80 with a donor plant comprisinga desired trait, selecting an F1 progeny plant comprising the desiredtrait, and backcrossing the selected F1 progeny plant to a plant ofsoybean variety 95M80. This method may further comprise the step ofobtaining a molecular marker profile of soybean variety 95M80 and usingthe molecular marker profile to select for a progeny plant with thedesired trait and the molecular marker profile of 95M80. In oneembodiment the desired trait is a mutant gene or transgene present inthe donor parent.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. 95M80 is suitable for use in a recurrentselection program. The method entails individual plants crosspollinating with each other to form progeny. The progeny are grown andthe superior progeny selected by any number of selection methods, whichinclude individual plant, half-sib progeny, full-sib progeny and selfedprogeny. The selected progeny are cross pollinated with each other toform progeny for another population. This population is planted andagain superior plants are selected to cross pollinate with each other.Recurrent selection is a cyclical process and therefore can be repeatedas many times as desired. The objective of recurrent selection is toimprove the traits of a population. The improved population can then beused as a source of breeding material to obtain new varieties forcommercial or breeding use, including the production of a syntheticcultivar. A synthetic cultivar is the resultant progeny formed by theintercrossing of several selected varieties.

Mass selection is a useful technique when used in conjunction withmolecular marker enhanced selection. In mass selection seeds fromindividuals are selected based on phenotype or genotype. These selectedseeds are then bulked and used to grow the next generation. Bulkselection requires growing a population of plants in a bulk plot,allowing the plants to self-pollinate, harvesting the seed in bulk andthen using a sample of the seed harvested in bulk to plant the nextgeneration. Also, instead of self pollination, directed pollinationcould be used as part of the breeding program.

Mutation Breeding

Mutation breeding is another method of introducing new traits intosoybean variety 95M80. Mutations that occur spontaneously or areartificially induced can be useful sources of variability for a plantbreeder. The goal of artificial mutagenesis is to increase the rate ofmutation for a desired characteristic. Mutation rates can be increasedby many different means including temperature, long-term seed storage,tissue culture conditions, radiation; such as X-rays, Gamma rays (e.g.cobalt 60 or cesium 137), neutrons, (product of nuclear fission byuranium 235 in an atomic reactor), Beta radiation (emitted fromradioisotopes such as phosphorus 32 or carbon 14), or ultravioletradiation (preferably from 2500 to 2900 nm), or chemical mutagens (suchas base analogues (5-bromo-uracil), related compounds (8-ethoxycaffeine), antibiotics (streptonigrin), alkylating agents (sulfurmustards, nitrogen mustards, epoxides, ethylenamines, sulfates,sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, oracridines. Once a desired trait is observed through mutagenesis thetrait may then be incorporated into existing germplasm by traditionalbreeding techniques. Details of mutation breeding can be found in“Principals of Cultivar Development” Fehr, 1993 Macmillan PublishingCompany the disclosure of which is incorporated herein by reference. Inaddition, mutations created in other soybean plants may be used toproduce a backcross conversion of 95M80 that comprises such mutation.

Breeding with Molecular Markers

Molecular markers, which includes markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs) and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods utilizing 95M80.

Isozyme Electrophoresis and RFLPs have been widely used to determinegenetic composition. Shoemaker and Olsen, ((1993) Molecular Linkage Mapof Soybean (Glycine max L. Merr.). p. 6.131–6.138. In S. J. O'Brien(ed.) Genetic Maps: Locus Maps of Complex Genomes. Cold Spring HarborLaboratory Press. Cold Spring Harbor, N.Y.), developed a moleculargenetic linkage map that consisted of 25 linkage groups with about 365RFLP, 11 RAPD (random amplified polymorphic DNA), three classicalmarkers, and four isozyme loci. See also, Shoemaker R. C. 1994 RFLP Mapof Soybean. P. 299–309 In R. L. Phillips and I. K. Vasil (ed.) DNA-basedmarkers in plants. Kluwer Academic Press Dordrecht, the Netherlands.

SSR technology is currently the most efficient and practical markertechnology; more marker loci can be routinely used and more alleles permarker locus can be found using SSRs in comparison to RFLPs. For exampleDiwan and Cregan, described a highly polymorphic microsatellite loci insoybean with as many as 26 alleles. (Diwan, N., and P. B. Cregan 1997Automated sizing of fluorescent-labeled simple sequence repeat (SSR)markers to assay genetic variation in Soybean Theor. Appl. Genet.95:220–225.) Single Nucleotide Polymorphisms may also be used toidentify the unique genetic composition of the invention and progenyvarieties retaining that unique genetic composition. Various molecularmarker techniques may be used in combination to enhance overallresolution.

Soybean DNA molecular marker linkage maps have been rapidly constructedand widely implemented in genetic studies. One such study is describedin Cregan et. al, “An Integrated Genetic Linkage Map of the SoybeanGenome” Crop Science 39:1464–1490 (1999). Sequences and PCR conditionsof SSR Loci in Soybean as well as the most current genetic map may befound in Soybase on the world wide web.

One use of molecular markers is Quantitative Trait Loci (QTL) mapping.QTL mapping is the use of markers, which are known to be closely linkedto alleles that have measurable effects on a quantitative trait.Selection in the breeding process is based upon the accumulation ofmarkers linked to the positive effecting alleles and/or the eliminationof the markers linked to the negative effecting alleles from the plant'sgenome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants. Itcan also be used to reduce the number of crosses back to the recurrentparent needed in a backcrossing program. The use of molecular markers inthe selection process is often called genetic marker enhanced selection.

Production of Double Haploids

The production of double haploids can also be used for the developmentof plants with a homozygous phenotype in the breeding program. Forexample, a soybean plant for which 95M80 is a parent can be used toproduce double haploid plants. Double haploids are produced by thedoubling of a set of chromosomes (1N) from a heterozygous plant toproduce a completely homozygous individual. For example, see Wan et al.,“Efficient Production of Doubled Haploid Plants Through ColchicineTreatment of Anther-Derived Maize Callus”, Theoretical and AppliedGenetics, 77:889–892,1989 and US2003/0005479. This can be advantageousbecause the process omits the generations of selfing needed to obtain ahomozygous plant from a heterozygous source.

Thus, an embodiment of this invention is a process for making asubstantially homozygous 95M80 progeny plant by producing or obtaining aseed from the cross of 95M80 and another soybean plant and applyingdouble haploid methods to the F1 seed or F1 plant or to any successivefilial generation. Based on studies in maize and currently beingconducted in soybean, such methods would decrease the number ofgenerations required to produce a variety with similar genetics orcharacteristics to 95M80. See Bernardo, R. and Kahler, A. L., Theor.Appl. Genet. 102:986–992, 2001.

In particular, a process of making seed retaining the molecular markerprofile of soybean variety 95M80 is contemplated, such processcomprising obtaining or producing F1 seed for which soybean variety95M80 is a parent, inducing doubled haploids to create progeny withoutthe occurrence of meiotic segregation, obtaining the molecular markerprofile of soybean variety 95M80, and selecting progeny that retain themolecular marker profile of 95M80.

Use of 95M80 in Tissue Culture

This invention is also directed to the use of variety 95M80 in tissueculture. Tissue culture of various tissues of soybeans and regenerationof plants therefrom is well known and widely published. For example,reference may be had to Komatsuda, T. et al., “Genotype X SucroseInteractions for Somatic Embryogenesis in Soybean,” Crop Sci. 31:333–337(1991); Stephens, P. A. et al., “Agronomic Evaluation ofTissue-Culture-Derived Soybean Plants,” Theor. Appl. Genet. (1991)82:633–635; Komatsuda, T. et al., “Maturation and Germination of SomaticEmbryos as Affected by Sucrose and Plant Growth Regulators in SoybeansGlycine gracilis Skvortz and Glycine max (L.) Merr.,” Plant Cell, Tissueand Organ Culture, 28:103–113 (1992); Dhir, S. et al., “Regeneration ofFertile Plants from Protoplasts of Soybean (Glycine max L. Merr.):Genotypic Differences in Culture Response,” Plant Cell Reports (1992)11:285–289; Pandey, P. et al., “Plant Regeneration from Leaf andHypocotyl Explants of Glycine wightii (W. and A.) VERDC. var.longicauda,” Japan J. Breed. 42:1–5 (1992); and Shetty, K., et al.,“Stimulation of In Vitro Shoot Organogenesis in Glycine max (Merrill.)by Allantoin and Amides,” Plant Science 81:(1992) 245–251; as well asU.S. Pat. No. 5,024,944, issued Jun. 18, 1991 to Collins et al. and U.S.Pat. No. 5,008,200, issued Apr. 16, 1991 to Ranch et al., thedisclosures of which are hereby incorporated herein in their entirety byreference. Thus, another aspect of this invention is to provide cellswhich upon growth and differentiation produce soybean plants having thephysiological and morphological characteristics of soybean variety95M80.

Progeny Plants

All plants produced by the use of the methods described herein and thatretain the unique genetic or trait combinations of 95M80 are within thescope of the invention. Progeny of the breeding methods described hereinmay be characterized in any number of ways, such as by traits retainedin the progeny, pedigree and/or molecular markers. Combinations of thesemethods of characterization may be used.

Breeders of ordinary skill in the art have developed the concept of an“essentially derived variety”, which is defined in 7 U.S.C. § 2104(a)(3)of the Plant Variety Protection Act and is hereby incorporated byreference. Varieties and plants that are essentially derived from 95M80are within the scope of the invention.

Pedigree is a method used by breeders of ordinary skill in the art todescribe the varieties. Varieties that are more closely related bypedigree are likely to share common genotypes and combinations ofphenotypic characteristics. All breeders of ordinary skill in the artmaintain pedigree records of their breeding programs. These pedigreerecords contain a detailed description of the breeding process,including a listing of all parental varieties used in the breedingprocess and information on how such variety was used. One embodiment ofthis invention is progeny plants and parts thereof with at least oneancestor that is 95M80, and more specifically, where the pedigree of theprogeny includes 1, 2, 3, 4, and/or 5 or less breeding crosses to asoybean plant other than 95M80 or a plant that has 95M80 as a parent orother progenitor. A breeder of ordinary skill in the art would know if95M80 were used in the development of a progeny variety, and would alsoknow how many crosses to a variety other than 95M80 or variety with95M80 as a parent or other progenitor were made in the development ofany progeny variety.

Molecular markers also provide a means by which those of ordinary skillin the art characterize the similarity or differences of two varieties.Using the breeding methods described herein, one can develop individualplants, plant cells, and populations of plants that retain at least 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%,68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%,82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or 99.5%, genetic contribution from soybean variety95M80, as measured by either percent identity or percent similarity. Inpedigree analysis the percentage genetic contribution may not beactually known, but on average 50% of the starting germplasm would beexpected to be passed to the progeny variety after one cross to anothervariety, 25% after another cross to a different variety, and so on. Withbackcrossing, the expected contribution of 95M80 after 2, 3, 4 and 5doses (or 1, 2, 3 and 4 backcrosses) would be 75%, 87.5%, 93.75% and96.875% respectively. Actual genetic contribution may be much higherthan the genetic contribution expected by pedigree, especially ifmolecular markers are used in selection. Molecular markers could also beused to confirm and/or determine the pedigree of the progeny variety.

Traits are also used by those of ordinary skill in the art tocharacterize progeny. Traits are commonly evaluated at a significancelevel, such as a 1%, 5% or 10% significance level, when measured inplants grown in the same environmental conditions. For example, abackcross conversion of 95M80 may be characterized as having the samemorphological and physiological traits as 95M80. The traits used forcomparison may be any or all of the traits shown in Table 1 or Table 2.Environmental conditions should be appropriate for the traits or traitsbeing evaluated. Sufficient selection pressure should be present foroptimum measurement of traits of interest such as herbicide, insect ordisease resistance. Similarly, an introgressed trait conversion of 95M80for resistance, such as herbicide resistance, should not be compared to95M80 in the presence of the herbicide when comparing non-resistancerelated traits such as plant height and yield.

The population of plants produced at each and any cycle of breeding isalso an embodiment of the invention, and on average each such populationwould predictably consist of plants containing approximately 50% of itsgenes from variety 95M80 in the first breeding cycle, 25% of its genesfrom variety 95M80 in the second breeding cycle, 12.5% of its genes fromvariety 95M80 in the third breeding cycle, 6.25% in the fourth breedingcycle, and so on. However, in each case the use of variety 95M80provides a benefit, because linkage groups of 95M80 are retained in theprogeny varieties. Specifically, an embodiment of the invention is aprocess for making a population of 95M80 progeny plants comprisingobtaining or producing a first generation progeny seed comprising theplant of 95M80 as a parent, growing said first generation progeny seedto produce first generation plants, obtaining self or sib pollinatedseed from said first generation plants, and growing the self or sibpollinated seed to obtain a population of 95M80 progeny plants.

The population of 95M80 progeny soybean plants produced by this methodwill retain the expected genetic contribution of 95M80 described above.A variety selected from the population of 95M80 progeny plants producedby this method is an embodiment, and such variety may be furthercharacterized by its molecular marker identity or similarity to 95M80.

In this manner, the invention encompasses a process for making asubstantially homozygous 95M80 progeny plant comprising the steps ofobtaining or producing a first generation progeny seed wherein a parentof said first generation progeny seed is a plant of variety 95M80,growing said first generation progeny seed to produce a first generationplant and obtaining self or sib pollinated seed from said firstgeneration soybean plant, and producing successive filial generations toobtain a substantially homozygous 95M80 progeny plant. Also anembodiment of this invention is the substantially homozygous 95M80progeny plant produced by this method.

Deposits

Applicant(s) have made a deposit of at least 2500 seeds of SoybeanVariety 95M80 with the American Type Culture Collection (ATCC),Manassas, Va. 20110, USA, ATCC Deposit No. PTA-5779. The seeds depositedwith the ATCC on Jan. 15, 2004 were taken from the deposit maintained byPioneer Hi-Bred International, Inc., 7250 NW 62nd Avenue, Johnston, Iowa50131 since prior to the filing date of this application. Access to thisdeposit will be available during the pendency of the application to theCommissioner of Patents and Trademarks and persons determined by theCommissioner to be entitled thereto upon request. Upon allowance of anyclaims in the application, the Applicant(s) will make the depositavailable to the public pursuant to 37 CFR 1.808. This deposit ofSoybean Variety 95M80 will be maintained in the ATCC depository, whichis a public depository, for a period of 30 years, or 5 years after themost recent request, or for the enforceable life of the patent,whichever is longer, and will be replaced if it becomes nonviable duringthat period. Additionally, Applicant(s) have or will satisfy all therequirements of 37 C.F.R. §§1.801–1.809, including providing anindication of the viability of the sample upon deposit. Applicant(s)have no authority to waive any restrictions imposed by law on thetransfer of biological material or its transportation in commerce.Applicant(s) do not waive any infringement of their rights granted underthis patent or under the Plant Variety Protection Act (7 USC 2321 etseq.). U.S. Plant Variety Protection of Soybean Variety 95M80 has issuedunder PVP certificate number 200400074.

All publications, patents and patent applications mentioned in thespecification are indicative of the level of those skilled in the art towhich this invention pertains. All such publications, patents and patentapplications are incorporated by reference herein for the purpose citedto the same extent as if each was specifically and individuallyindicated to be incorporated by reference herein.

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding. Asis readily apparent to one skilled in the art, the foregoing are onlysome of the methods and compositions that illustrate the embodiments ofthe foregoing invention. It will be apparent to those of ordinary skillin the art that variations, changes, modifications and alterations maybe applied to the compositions and/or methods described herein withoutdeparting from the true spirit, concept and scope of the invention.

1. Seed of soybean variety 95M80, representative seed of said soybeanvariety 95M80 having been deposited under ATCC Accession No. PTA-5779.2. A soybean plant, or a part thereof, produced by growing the seed ofclaim
 1. 3. The soybean plant part of claim 2, wherein said part ispollen.
 4. The soybean plant part of claim 2, wherein said part is anovule.
 5. A tissue culture of protoplasts or regenerable cells from theplant of claim
 2. 6. The tissue culture according to claim 5, whereincells or protoplasts of the tissue culture are obtained from planttissue selected from the group consisting of: leaf, pollen, cotyledon,hypocotyl, embryos, root, pod, flower, shoot and stem.
 7. A soybeanplant regenerated from the tissue culture of claim 5 and having all themorphological and physiological characteristics of soybean variety95M80, representative seed of said soybean variety 95M80 having beendeposited under ATCC Accession No. PTA-5779.
 8. A method for producing aprogeny soybean plant wherein the method comprises crossing the plant ofclaim 2 with a different soybean plant, harvesting the resultant soybeanseed, and growing a progeny soybean plant.